Research Institute for Biomacromolecules, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)

JINA SHIN

Research Institute for Biomacromolecules, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)

Jeong Hye Sunwoo

Research Institute for Biomacromolecules, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)

Eun Soon Kim

Research Institute for Biomacromolecules, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)

Jae Yong Kim

Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)Research Institute for Biomacromolecules, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)

Hungwon Tchah

Ophthalmology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)Research Institute for Biomacromolecules, University of Ulsan College of Medicine, Asan Medical Center, Seoul, Korea (the Republic of)

Footnotes

Commercial RelationshipsJin Hyoung Park, National Research Foundation of Korea (F); Soon-Suk Kang, National Research Foundation of Korea (F); JINA SHIN, National Research Foundation of Korea (F); Jeong Hye Sunwoo, National Research Foundation of Korea (F); Eun Soon Kim, National Research Foundation of Korea (F); Jae Yong Kim, National Research Foundation of Korea (F); Hungwon Tchah, National Research Foundation of Korea (F)

Purpose :
To examine the effects of nerve growth factor (NGF) on apoptosis, inflammation and epithelial-mesenchymal transition (EMT) in the diabetic lens.

Methods :
To investigate the effects of NGF on glucose-induced apoptosis, we stained human lens epithelial cells (HLECs) using fluorescein isothiocyanate (FITC)-labeled annexin-V and propidium iodide (PI), and measured expression of cleaved caspase-3 and the Bcl-2 by western blot. Moreover, to examine the effects of NGF on inflammation, we quantified the expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) using multiplex cytokine analysis, and analyzed nuclear factor-κB (NF-κB) activation and NF-κB inhibitor α (IκB-α) degradation using western blot analysis. And, to investigate the effects of NGF on EMT, we measured expression of α-smooth muscle actin (α-SMA) and vimentin. To investigate the effects in vivo, we induced diabetes in male Sprague–Dawley rats using streptozotocin. The rats were divided into 3 groups: “Control”, “Diabetic control”, and “Diabetic NGF”; topical NGF was applied three times daily for 8 weeks. We used immunohistochemistry to identify cleaved caspase-3, IL-1β and α-SMA.

Results :
In HLECs, high glucose concentration (≥ 25 mM) led to apoptosis, the release of inflammatory cytokines and EMT. NGF markedly reduced Annexin-PI-positive cells, and levels of cleaved caspase-3, IL-1β, TNF-α, α-SMA and vimentin and increased Bcl-2 expression. In the diabetic rat lens, apoptosis, inflammation and EMT were enhanced, as were levels of cleaved caspase-3, IL-1 β and α-SMA. These responses were markedly reduced by NGF.

Conclusions :
Apoptosis, inflammation and EMT are enhanced in the diabetic lens; NGF attenuates these responses—both in vivo and in vitro. Therefore, NGF therapy may represent a novel approach for the prevention and management of diabetic cataract.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.